�ber eine spezifische Anf�rbung der Pollenschl�uche im Griffel und die Zahl der Kallosepfropfen nach Selbstung und Fremdung

Linskens, H. F.; Esser, Karl

doi:10.1007/bf00629340

Cited by 262 publications

(138 citation statements)

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“…A high intracellular level of lily pollen invertase may have significance in the hydrolysis of endogenous sucrose during pollen germination and also that of sugars taken up from external nutritional sources. It has been shown that sugars are absorbed from stylar tissue by germinating pollen of L. regale and L. henryi (16). It is of significance that in L. auratum, the species used in our studies, sucrose is the major soluble sugar in mature pollen (21).…”

Section: Resultsmentioning

confidence: 95%

Invertases of Lilium Pollen

Singh¹,

Knox²

1984

Plant Physiol.

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ITwo different forms of invertase are found in pollen of lily (Lilium anratum). One form is cytoplasmic (Invertase 1) and the other is bound to the pollen wall (Invertase 2). Invertase 1 has been partilly purifed and is a glycoprotein (apparent molecular weight, 450 kilodaltons) with a K, of 0.65 millimolar for sucrose. The two invertases differ in pH optimum and thermal stability. Invertases of lily pollen are ,@-fructofuranosidases which can hydrolyze sucrose but not melizitose. The mature pollen grains have enzyme activity in both cytoplasmic and wall fractions, and no increase in the activity of either occurs during germtion. The wall-bound enzyme could not be released by treatments with detergents or high salt concentrations.Invertase (EC 3.2.1.26) or ,B-fructofuranoside fructohydrolase hydrolyzes sucrose and related oligosaccharides. Sucrose represents the major soluble sugar present in the majority of angiosperm pollen grains and is also the most commonly detected sugar in natural substrates for pollen germination (25). Germinating pollen grains show an extremely high rate of growth, i.e. 400 Am/h (13) and thus have a high demand for carbon skeletons required for pollen tube wall synthesis as well as a substrate for respiration. In vivo, the pollen tubes of lily grow on the surface of stigmatic and stylar canal secretion products. By means of labeling experiments, it has been shown that carbohydrate material of the secretion product of styles is used by growing pollen tubes for tube wall synthesis (17). Although invertase was among the first enzyme activities recognized in pollen, no detailed characterization of this pollen enzyme has been attempted. In the present study, we report the purification and properties of invertase from the pollen of Lilium auratum and discuss its functional significance. 12,000g for 10 min, and the supernatant was dialyzed for 20 h at 2C against extraction buffer and the dialyzed extracts were then used as a crude extract for enzyme assays. The wall pellets were again washed with extraction buffer three times and the washings were discarded. The resulting pellet was then suspended in 50 mM citrate-phosphate buffer (pH 6.0), and the suspension was used as a source of wall-bound invertase. Aliquots of wall fraction were assayed for hexokinase activity (11) to ascertain the absence of contaminant cytoplasmic proteins. The enzymic activity of soluble invertase was assayed as follows: a 0.05-ml sample of invertase preparation was added to 0.45 ml of 0.1 M acetate buffer (pH 4.8) which contained sucrose at a final concentration of 100 mm. The mixture was incubated for 20 min at 35C. The reactions were terminated by placing the reaction tubes in a boiling water bath for 5 min. After cooling the tubes to 35C, glucose released in the reaction mixture was estimated by a colorimetric assay using glucose oxidase and peroxidase. The assay was carried out by adding 0.5 ml ofchromogen reagent (4) to 0.5 ml ofthe invertase reaction mixture. In some instances, the reducing sugars relea...

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Section: Resultsmentioning

confidence: 95%

Invertases of Lilium Pollen

Singh¹,

Knox²

1984

Plant Physiol.

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“…Pollen tube growth was allowed to take place for 18 hr at 250 in a moist chamber, then stigmas were treated with decolorized aniline-blue:glycerol 9:1. Under these conditions, pollen tube growth was observed by the fluorescent staining of callose in the pollen tube wall (11). For treatment, the stigmas were dipped in solutions of the agents for 5 Con-A-Sepharose.…”

Section: Methodsmentioning

confidence: 99%

Cell recognition in plants: Determinants of the stigma surface and their pollen interactions

Knox

Clarke

Harrison

et al. 1976

Proc. Natl. Acad. Sci. U.S.A.

123

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The interaction between compatible pollen grains and the female stigma of Gladiolus gandavensis has been used as a model system for investigation of cell recognition in plants. The Cells of virtually all eukaryotic organisms possess some capacity to distinguish self from not-self (1). Processes reflecting this recognition capacity have been extensively explored in animal systems (2), but plants have been largely neglected. However, recognition between compatible pollen and the female stigma in plants provides a model system for cell recognition which is genetically specified (3), and in which transfer of information has been demonstrated both for pollen, which carries the male gametes (4), and the stigma surface (5). In this communication we describe the recognition events in the self-compatible system of Gladiolus, whose pollen walls are known to contain a range of proteins in both exine and intine sites (6). We have investigated the molecular architecture of the external stigma surface with a view to demonstrating surface receptors. (7) showed that the dialyzed and dried extract contained 25% sodium deoxycholate and 17% MATERIALS AND METHODS

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“…Decolourised aniline blue (Linskens & Esser, 1957) was used to visualize callose in both squash and sectioned material. Squashes were prepared by exerting gentle pressure on excised stigmas placed between a coverslip and a glass slide.…”

Section: Light Microscopymentioning

confidence: 99%

Identification of pollen components regulating pollination‐specific responses in the stigmatic papillae of Brassica oleracea

Elleman

Dickinson

1996

New Phytologist

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SUMMARYWhen components of the mature pollen grain of Brassica oleracea are applied to the cutieularised surfaces of the stigmatic papilla, a number of changes can take place in the architecture of the subjacent cell wall. Treatment of the papillar surface with isolated pollen coating evokes a rapid and extensive expansion of the outer of the two stigmatic wall layers. This response occurs within 4 h but is restricted to those regions where no callose is formed and where the coating exhibits a characteristic increase in electron opacity. This study establishes that the coating alone is responsible for initiating the expansion of the outer wall, which is considered to be an essential step preceding penetration ofthe stigma surface by the pollen tube. Vesicle-like inclusions, some staining intensely, occasionally occur in regions of the wall expanded hy isolated coating, sometimes fusing to form a subcuticular 'boundary layer'. These structures are not observed under compatible grains m vivo and their presence is regarded as artefactual. Howe\'er, close examination of the plasma membrane at sites beneath areas of expanded cell wall reveals membrane-bound structures resembling the vesicles being generated by the cytoplasm and moving into the apoplast. These data indicate that signals from the coating initiate the loosening of the wall matrix by stinnulating an unusual form of localized secretion which appears to be an essential prerequisite for stigmatic penetration.Isolated pollen coating contaminated with fragments of the grain itself engender a very different response, including the formation of densely-staining vesicles accompanied by extensive accumulations of callose; wall expansion never occurs under these circumstances. Self-pollinations in B. oleracea are frequently accompanied by the synthesis of stigmatic callose, the presence of which has, in the past, been interpreted as forming a struetural barrier to incompatible pollen tubes. However, callose elicited both by self-pollinations and coating supplemented by killed grains is independent of protein synthesis and occurs in the presenee of the phosphatase inhibitor, okadaic acid. Since both these inhibitors also overcome self-ineompatibility (SI) in B. oleracea, these data strongly suggest that callose synthesis is unrelated to the operation of SI, and it is proposed that callose, when formed, is elicited by molecules released from the necrotic pollen protoplast. Interestingly, isolated pollen coating of self-and cross-genotypes does not readily elicit callose.The significance of these ehanges in the stigmatic wall is discussed in the perspective of current views on pollination and self-incompatibility in Brassica.

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�ber eine spezifische Anf�rbung der Pollenschl�uche im Griffel und die Zahl der Kallosepfropfen nach Selbstung und Fremdung

Cited by 262 publications

References 3 publications

Invertases of Lilium Pollen

Invertases of Lilium Pollen

Cell recognition in plants: Determinants of the stigma surface and their pollen interactions

Identification of pollen components regulating pollination‐specific responses in the stigmatic papillae of Brassica oleracea

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